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HTS001M ChemiSCREEN™ Membrane Preparation Recombinant Human CXCR1 Chemokine Receptor

Human CXCR1 / IL-8a GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Host Cell: Chem-1 Uni Prot Number: P25024 Entrez Gene Number: NM_000634.2
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HTS001M
200 units  
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Overview

Replacement Information

Key Spec Table

SpeciesHost CellsProtein Target
Human Chem-1CXCR1 / IL-8a
Description
Catalogue NumberHTS001M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human CXCR1 Chemokine Receptor
Overview: Full-length human CXCR1 cDNA
Background InformationCXCR1 (also known as IL-8RA) is a Gi/o-coupled GPCR expressed on neutrophils, CD8(+) T cells, and intestinal epithelial cells (Murphy, 1997; Sturm et al., 2005; Takata et al., 2004). CXCR1 binds specifically to the chemokine interleukin-8 (IL-8), whereas a related receptor, CXCR2, binds promiscuously to IL-8 and several other chemokines. CXCR1 has been proposed to be the primary regulator of neutrophil response to IL-8 in sepsis (Cummings et al., 1999). Antagonists of CXCR1 and CXCR2 have been developed and display efficacy in animal models of reperfusion injury (Bertini, et al., 2004; Kaneider et al., 2005). Millipore's CXCR1 membrane preparations are crude membrane preparations made from our proprietary stable recombinant cell lines to ensure high-level of GPCR surface expression; thus, they are ideal HTS tools for screening of antagonists of CXCR1 interaction with IL-8. The membrane preparations exhibit a Kd of 0.2 nM for [125I]-IL-8. With 5 μg/well CXCR1 Membrane Prep and 0.1 nM [125I]-IL-8, a greater than 5-fold signal-to-background ratio is obtained.
References
Product Information
FormatMembranes
PresentationLiquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA no preservatives.Packaging method: Membranes protein were adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.
Quality LevelMQ100
Applications
ApplicationHuman CXCR1 / IL-8a GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.
Key Applications
  • Radioligand binding assay
  • GTPγS binding
Application NotesRadioligand binding assay, and GTPgammaS binding.
Biological Information
Concentration1 mg/mL
Entrez Gene Number
Entrez Gene SummaryThe protein encoded by this gene is a member of the G-protein-coupled receptor family. This protein is a receptor for interleukin 8 (IL8). It binds to IL8 with high affinity, and transduces the signal through a G-protein activated second messenger system. Knockout studies in mice suggested that this protein inhibits embryonic oligodendrocyte precursor migration in developing spinal cord. This gene, IL8RB, a gene encoding another high affinity IL8 receptor, as well as IL8RBP, a pseudogene of IL8RB, form a gene cluster in a region mapped to chromosome 2q33-q36.
Gene Symbol
  • IL8RA
  • CMKAR1
  • CKR-1
  • C-C
  • C-C-CKR-1
  • CXCR-1
  • IL8RBA
  • IL8R1
  • CDw128a
  • CD128
  • CXCR1
  • CD181
GPCR ClassA
Host CellsChem-1
Protein TargetCXCR1 / IL-8a
Target Sub-FamilyChemokine
UniProt Number
UniProt SummaryFUNCTION: SwissProt: P25024 # Receptor to interleukin-8, which is a powerful neutrophils chemotactic factor. Binding of IL-8 to the receptor causes activation of neutrophils. This response is mediated via a G-protein that activate a phosphatidylinositol-calcium second messenger system. This receptor binds to IL-8 with a high affinity and to MGSA (GRO) with a low affinity.
SIZE: 350 amino acids; 39791 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
SIMILARITY: SwissProt: P25024 ## Belongs to the G-protein coupled receptor 1 family.
Incubation ConditionsRECOMMENDED ASSAY CONDITIONS: Membranes are mixed with radioactive ligand and unlabeled competitor (see Figures 1 and 2 for concentrations tested) in binding buffer in a nonbinding 96-well plate, and incubated for 1-2 h. Prior to filtration, an FC 96-well harvest plate (Millipore cat. # MAHF C1H) is coated with 0.33% polyethyleneimine for 30 min, then washed with 50mM HEPES, pH 7.4, 0.5% BSA. Binding reaction is transferred to the filter plate, and washed 3 times (1 mL per well per wash) with Wash Buffer. The plate is dried and counted.

Binding buffer: 50 mM Hepes, pH 7.4, 5 mM MgCl2, 1 mM CaCl2, 0.2% BSA, filtered and stored at 4°C

Radioligand: [125I] Interleukin-8 (Perkin Elmer #NEX277)

Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.

One vial contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 5-fold signal:background with 125I-labeled IL-8 at 0.1 nM
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Table 1. Signal:background and specific binding values obtained in a competition binding assay with varying amounts of CXCR1 Receptor membrane preparation in a dose response curve with unlabeled IL-8.
Product Usage Statements
5 µg/well 10 µg/well
Signal:Background 14.8 12.8
Specific Binding (cpm) 8965 9624


SPECIFICATIONS: 1 unit = 5 μg

Bmax: 3.7 pmol/mg;

Kd: 0.2 nM Usage Statement
  • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage ConditionsMaintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
Packaging Information
Material Size200 units
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number GTIN
HTS001M 04053252283055

Documentation

Protocols

Title
Assembly of a Competition Binding Curve

ChemiSCREEN™ Membrane Preparation Recombinant Human CXCR1 Chemokine Receptor SDS

Title

Safety Data Sheet (SDS) 

ChemiSCREEN™ Membrane Preparation Recombinant Human CXCR1 Chemokine Receptor Certificates of Analysis

TitleLot Number
CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN CXCR1 RECEPTOR - 2218176 2218176

References

Reference overviewPub Med ID
Reversing systemic inflammatory response syndrome with chemokine receptor pepducins.
Kaneider NC et al.
Nat Med., 11(6):661-5 (2005)  2005

15880119 15880119
Noncompetitive allosteric inhibitors of the inflammatory chemokine receptors CXCR1 and CXCR2: prevention of reperfusion injury.
Bertini R et al.
Proc. Natl. Acad. Sci. U S A., 101(32):11791-6 (2004)  2004

15282370 15282370
Cutting edge: expression of chemokine receptor CXCR1 on human effector CD8+ T cells.
Takata H et al.
J. Immunol., 173(4):2231-5 (2004)  2004

15294933 15294933
Expression and function of the chemokine receptors CXCR1 and CXCR2 in sepsis.
Cummings CJ et al.
J. Immunol., 162(4):2341-6 (1999)  1999

9973513 9973513

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Life Science Research > Proteins and Enzymes > GPCR Membrane Preparations